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Nat:“睡得香睡得好”?神经元内信号级联 睡眠稳态调控

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发表于 2022-12-17 13:38:08 | 显示全部楼层 |阅读模式
本帖最后由 顾汉现 于 2022-12-17 13:47 编辑

Nature | 如何“睡得香睡得好”?神经元内信号级联说了算  睡眠稳态调控

BioArt

2022/12/16

论文
论文标题:Kinase signalling in excitatory neurons regulates sleep quantity and depth
作者:Kim, Staci J., Hotta-Hirashima, Noriko, Asano, Fuyuki, Kitazono, Tomohiro, Iwasaki, Kanako, Nakata, Shinya, Komiya, Haruna, Asama, Nodoka, Matsuoka, Taeko, Fujiyama, Tomoyuki, Ikkyu, Aya, Kakizaki, Miyo, Kanno, Satomi, Choi, Jinhwan, Kumar, Deependra, Tsukamoto, Takumi, Elhosainy, Asmaa, Mizuno, Seiya, Miyazaki, Shinichi, Tsuneoka, Yousuke, Sugiyama, Fumihiro, Takahashi, Satoru, Hayashi, Yu, Muratani, Masafumi, Liu, Qinghua, Miyoshi, Chika, Yanagisawa, Masashi, Funato, Hiromasa

期刊:Nature
发表时间:2022/12/07
数字识别码:10.1038/s41586-022-05450-1
摘要:Progress has been made in the elucidation of sleep and wakefulness regulation at the neurocircuit level1,2. However, the intracellular signalling pathways that regulate sleep and the neuron groups in which these intracellular mechanisms work remain largely unknown. Here, using a forward genetics approach in mice, we identify histone deacetylase 4 (HDAC4) as a sleep-regulating molecule. Haploinsufficiency of Hdac4, a substrate of salt-inducible kinase 3 (SIK3)3, increased sleep. By contrast, mice that lacked SIK3 or its upstream kinase LKB1 in neurons or with a Hdac4S245A mutation that confers resistance to phosphorylation by SIK3 showed decreased sleep. These findings indicate that LKB1–SIK3–HDAC4 constitute a signalling cascade that regulates sleep and wakefulness. We also performed targeted manipulation of SIK3 and HDAC4 in specific neurons and brain regions. This showed that SIK3 signalling in excitatory neurons located in the cerebral cortex and the hypothalamus positively regulates EEG delta power during non-rapid eye movement sleep (NREMS) and NREMS amount, respectively. A subset of transcripts biased towards synaptic functions was commonly regulated in cortical glutamatergic neurons through the expression of a gain-of-function allele of Sik3 and through sleep deprivation. These findings suggest that NREMS quantity and depth are regulated by distinct groups of excitatory neurons through common intracellular signals. This study provides a basis for linking intracellular events and circuit-level mechanisms that control NREMS.

所属学科:
神经科学
撰文 | 亦


近年来,睡眠和觉醒的调控在神经环路水平的阐释取得了很大进步【1,2】,一些神经元群体被报道参与睡眠稳态调控【3-5】,但负责调控的细胞信号通路和参与的神经元群体很大程度上还不为人知。SIK3是睡眠稳态调节中关键的组分,它在神经元中的表达会增加睡眠数量,导致非快速眼动睡眠(non-rapid eye movement sleep, NREMS)期的脑电图(electroencephalogram, EEG)δ电流增强【6】,它的剪切突变Sik3也会有类似效果,其残基S551对于睡眠增加也很关键,但这一通路的上下游分子途径还有待阐明。

2022年12月7日,来自日本筑波大学国际综合睡眠医学研究所的Masashi Yanagisawa和Hiromasa Funato在Nature上发表了题为Kinase signalling in excitatory neurons regulates sleep quantity and depth的文章,鉴定出了睡眠调控分子——组蛋白酶脱乙酰酶4(HDAC4)和调节睡眠及觉醒状态的信号级联LKB1–SIK3–HDAC4,揭示了大脑皮层和下丘脑兴奋性神经元中SIK3信号通路在NREMS期的调节作用(另一篇背靠背的类似工作详见:Nature背靠背|刘清华等揭开睡眠量转录调控之谜)。



通过对随机突变的C57B/6J雄性小鼠进行筛选,作者建立了一个长睡眠谱系Sleepy2 (Slp2),其N2代的清醒时间显著减少。外显子测序发现其具有Hdac4剪切位点突变,会产生较短的Hdac4转录本,且HDAC4的数量显著减少。作者又建立了一个带有同样剪切受体(splice acceptor, SA)突变的Hdac4修饰小鼠,纯合的小鼠除了Hdac4转录本异常和HDAC4蛋白减少外,还表现出生长缓慢,而杂合的Hdac4SA表现正常,清醒时间减少,NREMS数量增加,EEG δ电流增强,以上说明Hdac4的抑制促进了NREMS。进一步地,作者分析了class IIa其他HDACs,发现Hdac5SA小鼠NREMS数量和深度也增加,而Hdac7SA和Hdac9SA则没有影响。

HDAC4会经历核质穿梭,调节基因转录。作者观察到当睡眠需求较高时,核内HDAC4水平降低,这与杂合Hdac4缺陷中NREMS增加一致。之前的研究表明HDAC4的核定位和转录抑制效应受磷酸化调控,作者发现HDAC4是作为SIK3的底物来调节睡眠的,SIK3在S245位点磷酸化HDAC4,Sik3Slp/+小鼠S245位点的HDAC4磷酸化增加,核内HDAC4减少。以上说明SIK3对HDAC4的功能的增强依赖S245位点的磷酸化。进一步地,作者观察到杂合的Hdac4S245A小鼠NREMS时间减少,δ电流强度减弱,而用腺相关病毒在Sik3Slp小鼠中表达HDAC4时,嗜睡表型消失。此外,在线虫中,Sik3的类似物kin-29和Hdac4的类似物hda-4与嗜睡相关,与小鼠一样,hda-4作用在kin-29的下游调节睡眠行为。

SIK3的激酶活性依赖于T221被LKB1磷酸化,那么LKB1的缺失或T221的丙氨酸替代可能会减少NREMS。作者发现在它莫昔芬诱导的Syn1creERT;Lkb1flox/flox小鼠模型中,不依赖LKB1的SIK3(T221E)激酶使小鼠NREMS δ电流增强,说明LKB1作用于SIK3上游。杂合的Sik3Slp;T221A小鼠清醒时间更长,NREMS时间更短,睡眠参数与野生型小鼠更接近,说明Sik3Slp小鼠中NREMS数量增加和δ电流增强的特性依赖于完整的激酶活性。进一步地,作者发现HDAC4调控睡眠的现象存在于谷氨酸能神经元中,而非GABA神经元中,兴奋性神经元中SIK3信号通路通过作用于不同脑区调节NREMS质量和深度。

接下来,作者利用单核RNA-seq找到了谷氨酸神经元中差异表达的基因,AAV-Sik3Slp中有133上调基因,AAV-Sik3Slp中有32个下调的基因。睡眠剥夺主要影响谷氨酸神经元表达的基因,其中,2,685基因上调,394基因下调。Sik3Slp的表达和睡眠剥夺影响的基因富集突触相关条目,且都会导致Bdnf基因上调,此基因参与睡眠稳态调控。至此,作者已发现大脑皮层中谷氨酸能神经元中SIK3信号通路调控NREMS δ电流强度,具体表现为LKB1磷酸化SIK3,SIK3磷酸化HDAC4,促使其定位于细胞质,从而促进促睡眠基因的转录。


图1 SIK3-HDAC4在睡眠调控中的作用

综上,文章利用正向遗传学方法,结合靶向遗传学分析对小鼠进行研究,结果提示NREMS质量和深度是由不同组兴奋性神经元通过公共胞内信号调控的,为联系细胞事件与控制NREMS的神经环路机制打下基础。

制版人:十一


参考文献

1. Adamantidis, A. R., Gutierrez Herrera, C. & Gent, T. C. Oscillating circuitries in the sleeping brain. Nat. Rev. Neurosci. 20, 746–762 (2019).
2. Liu, D. & Dan, Y. A motor theory of sleep–wake control: arousal–action circuit. Annu. Rev. Neurosci. 42, 27–46 (2019).
3. Reichert, S., Pavón Arocas, O. & Rihel, J. The neuropeptide galanin is required for homeostatic rebound sleep following increased neuronal activity. Neuron 104, 370–384. e5 (2019).
4. Oikonomou, G. et al. The serotonergic Raphe promote sleep in zebrafish and mice. Neuron 103, 686–701.e8 (2019).
5. Ma, Y. et al. Galanin neurons unite sleep homeostasis and α2-adrenergic sedation. Curr. Biol. 29, 3315–3322.e3 (2019).
6. Iwasaki, K. et al. Induction of mutant Sik3Sleepy allele in neurons in late infancy increases sleep need. J. Neurosci. 41, 2733–2746 (2021).

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文章标签
睡眠
觉醒
神经环路
睡眠调控分子
组蛋白酶脱乙酰酶4(HDAC4)
LKB1–SIK3–HDAC4信号级联
下丘脑
兴奋性神经元
SIK3信号通路

https://www.nature.com/articles/s41586-022-05450-1

https://www.linkresearcher.com/t ... d-be00-8030ebac7750



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